1 / 25

Toxicovenomics 2015

Invited lecture presentation at the 18th World Congress of the International Society on Toxinology, University of Oxford, UK, SEptember 2015

lomonte
Download Presentation

Toxicovenomics 2015

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. ‘Toxicovenomics’ towards an integrative view of compositional and functional aspects of snake venoms Invited lecture presented at the 18th World Congress of the International Society on Toxinology, University of Oxford, UK, September 2015 Bruno Lomonte

  2. Proteomics and snake venom composition • impact of proteomic tools upon venom composition • diverse analytical strategies for protein identification • combination with other ‘-omic’ tools (transcriptomics, genomics, ...)

  3. Snake venom proteomes • growing number of venom proteomes • increasingly sensitive methods • study of ‘scarce’ venoms from ‘uncommon’ species Cumulative number of species with venom proteomes studied Year

  4. Viperidae ~ 329 Elapidae ~ 417 Colubridae ~ 840 S ~ 1586 Serpentes ~ 3,500 species Venom proteomes by family of snakes (~200) Elapidae Viperidae Colubridae

  5. Gel-based strategies 2D-GE approach MS/MS analysis Gel-free (LC) strategies nano-flow LC Snake venom proteomes • early studies aimed at a thorough cataloguing of proteins • essentially qualitative information • identification frequency ≠ protein abundance • diverse analytical strategies

  6. Venomics strategy + LC offline + 1D GE approach MS/MS analysis • A combined (LC-offline + 1D-gel) approach • estimates relative protein abundances • adds relevant biological significance to cataloguing Snake venomics: estimation of abundances

  7. relative abundances provide key information to medical and biological significance • two hipothetical venoms consisting of the same components (A, B, C)

  8. Proteomic profiles of the venoms of Costa Rican snakes Lomonte et al. (2014) J. Proteomics 105, 323

  9. Proteomic profiles of the venoms of Costa Rican snakes Lomonte et al. (2014) J. Proteomics 105, 323

  10. Genus-wide venomic profiling: Agkistrodon spp. A. c. phaeogaster A. c. contortrix A. p. leucostoma A. c. laticinctus A. c. mokasen A. p. piscivorus A. c. pictagaster A. p. conanti A. taylori A. b. howardgloydi A. b. bilineatus Lomonte et al. (2014) J. Proteomics 96, 103

  11. Further collaborative ‘venomics’ studies Crotalus s. culminatus Crotalus s. simus Porthidium lansbergii Micrurus mipartitus Bothrops ayerbei Micrurus dumerilii Bothrops punctatus Bothrops pirajai Bothrocophias campbelli

  12. Venomics Antivenomics Toxicovenomics catalogue, + abundance, + antigenicity catalogue, + abundance, + functionality catalogue, + abundance Evolving strategies in venom analysis by proteomic tools Informative value catalogue Timeline

  13. How to combine venomics with functional characterization? Toxicity

  14. Pelamis platura (yellow-bellied sea snake) Lomonte et al. (2014) J. Proteomics 103, 137

  15. Case studies Micrurus tener Mus musculus Conopsis lineata Bénard-Valle et al. (2014) Toxicon77, 6-15

  16. black mamba Dendroaspis polylepis Naja kaouthia Aipysurus laevis Laustsen et al. (2015) J. Proteomics119, 126; Lausten et al. (2015) Toxicon99, 23; Laustsen et al.(2015), Toxicon in press

  17. Dendroaspis polylepis Which are the main toxic components in mice?

  18. LD50 (mg) Peak components Toxicity score (%/LD50) n.d. n.p. / nucleosides n.d n.p. / nucleosides n.d n.p. / nucleosides 0.08 3FTx 45.0 0.46 3FTx / Kun (dendrotoxin) 21.7 0.53 3FTx / Kun (dendrotoxin) 23.8 0.28 3FTx / Kun (dendrotoxin) 29.6 5.25 Kun (dendrotoxin) 4.6 3.76 3FTx / Kun (dendrotoxin) 1.7 4.92 3FTx / Kun (dendrotoxin) 0.5 4.38 3FTx / Kun (dendrotoxin) 0.8 >1.5 3FTx / Kun (dendrotoxin) <0.5 >5 3FTx <1 >2.5 3FTx <1.5 >1 3FTx >2.5 Kun (dendrotoxin) <0.5 >7.5 3FTx <0.5 >1 NGF <1 >5 3FTx <0.5 ? atypical Kun (dendrotoxin) n.d. >1.15 MP <1 >1 PDE <1 >0.3 HYA <1.5 n.d. MP n.d. n.d. MP n.d.

  19. Aipysurus laevis Laustsen et al. (2015) Toxicon (in press)

  20. Absorbance 215 nm (mU) 1000 2000 3000 0 0 10 1 20 2 3 4 30 5 6 40 9 8 7 10 11 12 13 17 14 Time (min) 50 18 15 16 60 70 19 80 20 90 LD50 (mg) % Peak/ component Toxicity score (%/LD50) 1 3FTx 0.07 21.8 334 2 3FTx 0.18 0.6 3.3 3 3FTx 0.13 2.0 15.3 4 3FTx 0.28 0.9 3.2 5 PLA2 >0.3 0.3 <1 6 PLA2 >0.5 0.5 <1 7 unk >0.25 0.2 <1 8 unk >0.5 0.4 <1 9 PLA2 >10 9.6 <1 10 PLA2 >7.5 6.3 <1 11 PLA2 >20 19.4 <1 12 PLA2 >6 6.0 <1 13 PLA2 >10 11.0 <1.1 14 PLA2 >2.5 2.5 <1 15 PLA2 >0.3 0.3 <1 16 PLA2 >0.25 0.2 <1 17 PLA2 3.05 3.6 1.2 18 CCM >2.5 11.6 <5 19 CRiSP >0.5 2.7 <1 20 CRiSP n.t. 0.1 n.t.

  21. Toxicovenomics Recent studies have begun to explore the possibilities of combining the venom compositional data provided by 'venomics', with the toxicity screening of its components - a strategy that could be named 'toxicovenomics'

  22. Toxicovenomics The value of the combined information provided by 'venomics' may be further expanded by integrating it with a functional assessment of the various venom peptides/proteins, in terms of their toxicity and other bioactivities

  23. chromatography: denaturing of labile proteins • high chromatographic resolution under native conditions • in vitro surrogate methods for toxic activities/bioassays Challenges

  24. This integrative approach has the potential to disclose the most clinically-relevant toxins within a particular venom It may provide a broader view on the biological and medical significance of snake venoms Toxicovenomics

  25. Davinia Pla Libia Sanz Juan J. Calvete Andreas Laustsen Mahmood Sasa Julián Fernández José M. Gutiérrez Bruno Lomonte

More Related